Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Combined Treatment of Nonsteroidal Anti-inflammatory Drugs and Genistein Synergistically Induces Apoptosis via Induction of NAG-1 in Human Lung Adenocarcinoma A549 Cells

인간 A549 폐암세포에서 비스테로이드성 항염증제와 genistein의 복합처리에 의한 NAG-1 의존적 세포사멸 증진 효과

  • Kim, Cho-Hee (Department of Molecular Biology, College of Natural Sciences, and Research Institute of Genetic Engineering, Pusan National University) ;
  • Kim, Min-Young (Department of Molecular Biology, College of Natural Sciences, and Research Institute of Genetic Engineering, Pusan National University) ;
  • Lee, Su-Yeon (Department of Molecular Biology, College of Natural Sciences, and Research Institute of Genetic Engineering, Pusan National University) ;
  • Moon, Ji-Young (Department of Molecular Biology, College of Natural Sciences, and Research Institute of Genetic Engineering, Pusan National University) ;
  • Han, Song-Iy (Research Center for Resistant Cells, College of Medicine, Chosun University) ;
  • Park, Hye-Gyeong (Korea Nanobiotechnology Center, Pusan National University) ;
  • Kang, Ho-Sung (Department of Molecular Biology, College of Natural Sciences, and Research Institute of Genetic Engineering, Pusan National University)
  • 김초희 (부산대학교 자연과학대학 분자생물학과, 유전공학연구소) ;
  • 김민영 (부산대학교 자연과학대학 분자생물학과, 유전공학연구소) ;
  • 이수연 (부산대학교 자연과학대학 분자생물학과, 유전공학연구소) ;
  • 문지영 (부산대학교 자연과학대학 분자생물학과, 유전공학연구소) ;
  • 한송이 (조선대학교 의과대학 내성세포연구센터) ;
  • 박혜경 (한국나노바이오테크놀러지센터) ;
  • 강호성 (부산대학교 자연과학대학 분자생물학과, 유전공학연구소)
  • Published : 2009.08.30
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Abstract

A number of studies have demonstrated that the regular use of nonsteroidal anti-inflammatory drugs (NSAIDs) can reduce the risks of colorectal, oesophageal and lung cancers. NSAIDs have been shown to exert their anti-cancer effects through inducing apoptosis in cancer cells. The susceptibility of tumor cells to anti-tumor drug-induced apoptosis appears to depend on the balance between pro-apoptotic and anti-apoptotic programs such as nuclear factor kB (NF-kB), phosphatidylinositol 3-kinase (PI3K)-Akt/protein kinase B (PKB) and MEK1/2-ERK1/2 pathways. We examined the effects of pro-survival PI3K and ERK1/2 signal pathways on cell cycle arrest and apoptosis in response to NSAIDs including sulindac sulfide and NS398. We show that simultaneous inhibition of the Akt/PKB and ERK1/2 signal cascades could synergistically enhance the potential pro-apoptotic activities of sulindac sulfide and NS398. Similar enhancement was observed in cells treated with sulindac sulfide or NS398 and 100 ${\mu}$M genistein, an inhibitor of receptor tyrosine kinases (RTKs) that are upstream of PI3K and MEK1/2 signaling. We further demonstrate that NAG-1 is induced and plays a critical role(s) in apoptosis by NSAIDs-based combined treatment. In sum, our results show that combinatorialtreatment of sulindac sulfide or NS398 and genistein results in a highlysynergistic induction of apoptotic cell death to increase the chemopreventive effects of the NSAIDs, sulindac sulfide and NS398.

비스테로이드성 항염증약(nonsteroidal anti-inflammatory drugs; NSAIDs)은 항염 및 진통효과를 나타내며, 염증억제 외에 다양한 신호전달 분자를 통해 여러 가지 세포생리활성을 조절하며, 암세포에서는 세포사멸 유도를 통한 항암제 효과를 보이고 있다. 본 연구에서는 NSAIDs가 암세포사멸프로그램을 작동시키는데 있어 phosphatidyl inositol 3-kinase (PI3K)-Akt/protein kinase B (PKB) 그리고 MEK1/2-ERK1/2 신호 전달계과 같은 anti-apoptotic program이 NSAIDs의 효과를 경감시키는 것으로 예상하고, 이들 항세포사멸 프로그램을 억제하였을 경우, NSAIDs의 세포사멸 유도작용이 증가되는지 그 가능성을 조사하였다. 세포사멸은 Hoeschst 33342으로 핵응축과 핵 쪼개짐을 염색하여 확인하였다. Western blotting을 통해 단백질 발현과 역전사중합효소연쇄반응을 통해 mRNA 발현을 확인하였다. NSAIDs 처리와 동시에 PI3K-Akt/PKB와 MEK-ERK1/2 신호전달계의 억제제를 함께 처리했을 때, NSAIDs의 세포사멸유도작용이 증가함을 확인하였다. 또한 PI3K와 MEKl/2 신호전달계의 상위에 존재하는 receptor tyrosine kinases (RTKs)의 억제제인 genistein을 함께 처리하였을 때에도 유사한 효과가 나타남을 확인하였다. 그리고 이들 복합처리에 의해 NAG-1 발현이 증가하며 NAG-1 interference 하였을 경우 복합처리에 의한 세포사멸증진 효과가 사라짐을 확인하였다. 본 연구결과는 암세포에 활성화 되어 있는 세포생존프로그램을 제어하는 물질(genistein 혹은 LY294002+U0126)을 복합처방함으로써 NSAIDs의 항암작용을 증진시킬 수 있음을 보여준다.

Keywords

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